Isolation of functional Golgi-derived vesicles with a possible role in retrograde transport.
Secretory proteins enter the Golgi apparatus when transport vesicles fuse with the cis-side and exit in transport vesicles budding from the trans-side. Resident Golgi enzymes that have been transported in the cis-to-trans direction with the secretory flow must be recycled constantly by retrograde transport in the opposite direction. In this study, we describe the functional characterization of Golgi-derived transport vesicles that were isolated from tissue culture cells. We found that under the steady-state conditions of a living cell, a fraction of resident Golgi enzymes was found in vesicles that could be separated from cisternal membranes. These vesicles appeared to be depleted of secretory cargo. They were capable of binding to and fusion with isolated Golgi membranes, and after fusion their enzymatic contents most efficiently processed cargo that had just entered the Golgi apparatus. Those results indicate a possible role for these structures in recycling of Golgi enzymes in the Golgi stack.
Love
H D
HD
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-0146, USA.
Lin
C C
CC
Short
C S
CS
Ostermann
J
J
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001692:Biological Transport; D016466:CHO Cells; D002458:Cell Fractionation; D018699:Coated Vesicles / Q000378:metabolism; D020756:Coatomer Protein; D006224:Cricetinae; D003600:Cytosol / Q000378:metabolism; D006056:Golgi Apparatus / Q000201:enzymology / Q000378:metabolism*; D007425:Intracellular Membranes / Q000201:enzymology / Q000378:metabolism*; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000378:metabolism; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism; D015388:Organelles / Q000201:enzymology / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000378:metabolism
D020756:Coatomer Protein; C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011506:Proteins; D014759:Viral Envelope Proteins; D000255:Adenosine Triphosphate; D017351:N-Acetylglucosaminyltransferases